7 3 Vol. 7 No. 3 2016 3 Journal of Food Safety and Quality Mar., 2016 李莉, 曹进 *, 张庆生 (, 100050) 摘要 : 目的 方法 60%, Agilent ZORBAX SB-C 18 (4.6 mm 250 mm, 5 μm), 0.5%, 1.0 ml/min, 30, 237 nm 结果 5.0~100.0 g/ml (r=0.9999), 95%~101%, RSD 0.55%~1.96% 结论, 关键词 : ; ; Determination of geniposidic acid in herba cistanche by high performance liquid chromatography LI Li, CAO Jin *, ZHANG Qing-Sheng (National Institutes for Food and Drug Control, Beijing 100050, China) ABSTRACT: Objective To establish a method for the determination of the content of geniposidic acid in herba cistanche by high performance liquid chromatography (HPLC). Methods Samples were extracted ultrasonically with 60% methanol as the solvent. An Agilent ZORBAX SB-C 18 column (4.6 mm 250 mm, 5 μm) was used with the mobile phase consisted of methanol and 0.5% acetic acid. The flow rate was 1.0 ml/min, and the column temperature was set at 30. The UV detection wavelength was 237 nm. Results There was a good linearity when the concentration of geniposidic acid was 5.0~100.0 mg/ml (r=0.9999). The average recovery was 95%~101%, and the RSD was 0.55%~1.96%. There were differences between the 3 different sources of hebra cistanehe (Cistanche deserticola, Cistanche tubulosa and Cistanche salsa) in geniposidic acid content. Conclusion This method is simple and accurate, and it can be used for determination of geniposidic acid in herba cistanche. KEY WORDS: hebra cistanehe; geniposidic acid; high performance liquid chromatography 1 引言 (hebra cistanehe) (Cistanche deserticola Y.C. Ma) [1],,,,, [2], 2005 (Cistanche tubulosa (Schrenk) Wight), * 通讯作者 :,, E-mail: caojin@gmail.com *Corresponding author: CAO Jin, Researcher, National Institutes for Food and Drug Control, No. 2, Tiantanxili, Dongcheng District, Beijing 100050, China. E-mail: caojin@gmail.com
934 7 [3],, 2, (Cistanche salsa (C.A.Mey) G. Beck) [4] (geniposidic acid),,,, [5-8] ( ),,, [9-14], 3 ( ) 2 材料与方法 2.1 仪器与试剂 2695 ( 2998, Waters ); AL 204 ( METTLER ) XP 205 ( METTLER ); CR22GIII ( HITACHI ); KQ-3200DE ( ) (, 121276-200501); (, 121101-200402); ( ); (, : 11828-201102, 96%); (, Merk); (, ); 2.2 仪器条件 : Agilent ZORBAX SB-C 18 (4.6 mm 250 mm, 5 μm); : A:, B:0.5%,, 1; : 1.0 ml/min; : 30 ; : 10 μl; 237 nm 2.3 标准溶液的制备 10 mg 10 ml, 1.0 mg/ml 表 1 梯度洗脱比例 Table 1 Gradient elution ratio (min) A(%) B(%) 0 5 95 10 20 80 25 20 80 35 60 40 45 5 95 2.4 样品前处理 2 g, 60% 30 ml, 30 min, 60%,, 50 ml, 0.45 μm, 3 结果与讨论 3.1 前处理方法的优化 ( ), 3, 3,,, 60:40, (15 30 45 min), 30 min, 30 min 3.2 液相色谱条件的优化 3.2.1 检测波长的选择 190~400 nm, 237 nm, 237 nm,,, 3.2.2 流动相的选择, - -,,,,,,,, +0.5%, 1
3, : 935 1 Fig. 1 High performance liquid chromatograms of geniposidic acid a: ; b: a: Spectrum of geniposidic acid standard; b: Spectrum of sample 3.2.3 柱温的选择 20 30 40,, 30 3.3 方法学考察 3.3.1 线性关系考察, 60% 6, 5.0 10.0 20.0 30.0 40.0 100.0 μg/ml 10 μl,, (Y) (X, g/ml) (r) Y=7139.4X+947.02(r=0.9999) 5.0~100.0 g/ml 3.3.2 加标回收试验, 3 (125 1000 2500 g/g) ( 6 ), 2.4, 10 L,, 2 3.3.3 重复性 2.4 6, HPLC, RSD 1.02%, 3.3.4 精密度 5 μg/ml 100 μg/ml, 6,, RSD 0.14% 0.06%, 3.3.5 稳定性, 0 2 4 8 12 24 48 72 h, HPLC, RSD 0.75%(n=6), 72 h 3.3.6 检测限和定量限,, 3, 10, 15 μg/g 50 μg/g
936 7 表 2 回收率测定结果 (n=6) Table 2 The results of recoveries (n=6) ( g/g) ( g/g) ( g/g) (%) (%) RSD(%) 118.35 125 237.07 94.97 118.35 125 237.42 93.66 118.35 125 237.54 95.35 118.35 125 239.64 97.03 95.3 0.95 118.35 125 238.37 96.01 118.35 125 236.27 94.33 118.35 1000 1126.1 100.8 118.35 1000 1100.7 98.24 118.35 1000 1111.0 99.27 118.35 1000 1119.2 100.1 99.79 0.69 118.35 1000 1119.5 100.1 118.35 1000 1121.1 100.3 118.35 2500 2619.4 100.0 118.35 2500 2624.2 100.2 118.35 2500 2593.6 99.01 118.35 2500 2619.1 100.0 99.5 0.55 118.35 2500 2590.9 98.90 118.35 2500 2595.7 99.10 3.4 样品测定, 3 3, [15] 表 3 实际样品测定结果 Table 3 The content of geniposidic acid in samples ( g/g) RSD(%) 153.8 0.02 556.5 0.15 / / : / 4 结论,, 参考文献 [1]. ( )[M]. :, 2010. China Pharmacopoeia Committee. Chinese pharmacopoeia (2010 Edition) [M]. Beijing: Chinese Medical Science and Technology Press, 2010. [2],,,. [J]., 2004, 6(1): 57 62. Sheng JH, Zhai ZX, Yang TX, et al. Study on biology of parasitism in Cistanche [J]. Rev China Agric Sci Technol, 2004, 6(1): 57 62. [3],,,. [J]., 2012, 35(7): 878 880. Jia XG, Ni H, Zhang J, et al. Present situation of research and
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